补肾健脾方介导肠道稳态改善去卵巢大鼠的骨量丢失  

The Efficacy of a kidney-tonifying and spleen-strengthening prescription in mediating gastrointestinal homeostasis and mitigating bone loss in ovariectomized rats

作　　者：谭日威 戴新华 叶嘉威 招文华 尚奇[1,3] 陈桂锋[1,3] 周本根 梁德 沈耿杨[2] 程英雄 范琳燕 任辉 江晓兵 TAN Riwei;DAI Xinhua;YE Jiawei;ZHAO Wenhua;SHANG Qi;CHEN Guifeng;ZHOU Bengen;LIANG De;SHEN Gengyang;CHENG Yingxiong;FAN Linyan;REN Hui;JIANG Xiaobing(The First Clinical College of Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Third Clinical College,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州中医药大学第一临床医学院,广东广州510405 [2]广州医科大学附属第二医院,广东广州510260 [3]广州中医药大学岭南医学研究中心,广东广州510405 [4]广州中医药大学第三临床医学院,广东广州510405 [5]广州中医药大学第一附属医院,广东广州510405

出　　处：《中国骨质疏松杂志》2024年第12期1717-1723,共7页Chinese Journal of Osteoporosis

基　　金：广东省自然科学基金面上项目(2021A1515011463,2021A1515011247);广州市科技计划(2022010117612)。

摘　　要：目的 探讨肠道稳态在去卵巢(OVX)大鼠骨量丢失中的机制及补肾健脾方的防治作用。方法 18只4月龄的雌性未孕SD大鼠随机平均分为假手术(SHAM)组、去卵巢(OVX)组、补肾健脾方(BSJP)组,每组6只。造模5 d后,SHAM组及OVX组予生理盐水灌胃,BSJP组予补肾健脾方灌胃,连续灌胃12周后取材。大鼠股骨的皮质骨及松质骨采用Micro-CT进行分析,小肠及股骨组织HE染色后镜下观察形态,TRAP染色后对破骨细胞数量进行统计,Western Blot检测肠道屏障关键蛋白Cldn3、小肠及骨组织内的免疫细胞关键标志物TNF-α、IL-17A及趋化因子CCL20、CXCR3、骨组织内破骨标志蛋白C-FOS、DC-STAMP、CTSK表达情况。结果 与OVX组比较,BSJP组肠壁增厚,小肠绒毛结构破坏减轻,股骨内破骨活动减少,Micro-CT骨微结构扫描提示BMD均明显升高;肠道屏障标志蛋白Cldn3蛋白表达上升,免疫细胞关键标志物TNF-α、IL17A及趋化因子CCL20、CXCR3蛋白表达均下降,破骨标志蛋白C-FOS、DC-STAMP、CTSK表达均下降。结论 去卵巢(OVX)大鼠发生骨质疏松的病理基础是肠道稳态失衡,补肾健脾方介导肠道稳态改善去卵巢(OVX)大鼠的骨量丢失。Objective To explore the mechanism of intestinal homeostasis in bone loss in ovariectomized(OVX)rats and the preventive and therapeutic effects of kidney tonifying and spleen strengthening formula.Methods A cohort of eighteen 4-month-old female Sprague-Dawley(SD)rats,which were nulliparous,were randomly assigned into three groups:a sham-operated control group(SHAM),an ovariectomized group(OVX),and a group treated with the kidney-tonifying and spleen-strengthening prescription(BSJP),with each group comprising six subjects.Following a 5-day post-operative recovery period,the SHAM and OVX groups were administered saline via gavage,whereas the BSJP group received the prescribed formulation via the same route for a duration of 12 weeks prior to sample collection.Analyses included Micro-CT imaging of the femur to evaluate bone density and microarchitecture,histological examination with hematoxylin and eosin(HE)staining of the small intestine and femur tissues,enumeration of osteoclasts through tartrate-resistant acid phosphatase(TRAP)staining,and Western Blot analyses to quantify the expression levels of the intestinal barrier protein Claudin-3(Cldn3),pro-inflammatory cytokines TNF-αand IL17A,chemokines CCL20 and CXCR3,and osteoclast-associated proteins C-FOS,DC-STAMP,and Cathepsin K(CTSK)in both the intestinal and bone tissues.Results Comparative analysis between the OVX and BSJP groups revealed a significant thickening of the intestinal wall,attenuation of damage to the villous architecture of the small intestine,and a reduction in osteoclastic activity within the femur in the BSJP group.Micro-CT scans demonstrated a notable increment in bone mineral density(BMD).Elevated expression of the intestinal barrier protein Cldn3 was observed in conjunction with a decrease in the levels of pro-inflammatory markers TNF-α,IL17A and chemokines CCL20,CXCR3 as well as a reduction in osteoclast-associated proteins C-FOS,DC-STAMP,and CTSK.Conclusion Osteoporosis in ovariectomized(OVX)rats stems from a dysregulation of gastrointes

关 键 词：绝经后骨质疏松症 补肾健脾方 骨量丢失 肠道稳态 

分 类 号：R285.5[医药卫生—中药学]